Method Development for Multiplexing CGE-LIF Based Glycoanalysis

Proteomics Along the Adaptation of an Adherent MDCK Cell Line to Growth in Suspension

In cooperation with Prof. Scharfenberg at the Applied University Emden/Leer in Emden/Germany an adherent Madin-Darby canine kidney (MDCK) cell was adapted to growth in suspension. In animal cell culture growth in suspension and in chemically defined medium is desirable for many production processes. However, for vaccine production processes, a switch to growth in suspension is often challenging, as many viruses seem to depend on cell lines difficult to adapt. Furthermore, cell-specific productivity is often reduced after adaptation.

Figure 1: Proteomes of a MDCK cell line grown on a surface (green) and grown in suspension (red) are compared by two-dimensional differential gel electrophoresis (2-D DIGE).
Figure 2: Proteomes of a cell line not infected (Control Bioreactor, green) and infected (Infection Bioreactor, red) are compared by 2D-DIGE.

In this project, adherent growth versus growth in suspension for a MDCK cell line was investigated on the proteome level. Interestingly, significant changes on the protein level during adaptation of an adherent MDCK cell to growth in suspension could be identified. Differences in proteomic profile were detected during cell line adaptation as well as during exponential and stationary growth of the adherent and the suspension cell line. Most of the differentially expressed proteins were subsequently identified. The findings gave new insights into key characteristics of cell line adaptation to growth in suspension.

Publications

Kluge, S.; Benndorf, D.; Genzel, Y.; Scharfenberg, K.; Rapp, E.; Reichl, U.: Monitoring changes in proteome during stepwise adaptation of a MDCK cell line from adherence to growth in suspension. Vaccine 33 (35), pp. 4269 - 4280 (2015)
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